KR20090057074A - Inhibition of angiogenesis and treatment of diseases related to angiogenesis - Google Patents

Abstract

The present invention provides a method of inhibiting angiogenesis in an individual by administering a composition (eg, a protein containing composition) comprising colchicine or thiocolhitine dimers. The composition is an amount effective to inhibit angiogenesis but in some embodiments is insufficient to cause significant cytotoxicity in the subject. The methods described herein are useful for treating angiogenesis related diseases such as senile macular degeneration, diabetic retinopathy, rheumatoid arthritis, psoriasis and cancer.

Description

{METHODS OF INHIBITING ANGIOGENESIS AND TREATING ANGIOGENESIS-ASSOCIATED DISEASES}

The present application relates to methods of inhibiting angiogenesis and treating angiogenesis related diseases. In particular, the present application relates to methods of inhibiting angiogenesis and treating angiogenesis related diseases by administering an effective amount of a composition comprising colchicine or thiocolhitine dimers.

Angiogenesis is a highly regulated biological process that forms new blood vessels. Uncontrolled angiogenesis can cause many diseases. One such disease is senile macular degeneration (“AMD”), which is characterized by the ingress of new blood vessels into various structures of the eye such as macular and retinal pigment epithelium. Another disease associated with angiogenesis is rheumatoid arthritis, in which angiogenesis occurs in the blood vessels of the synovial joint. In addition to forming new vascular networks, endothelial cells release reactive oxygen species and factors that cause pannus growth and cartilage destruction. Uncontrolled angiogenesis is also associated with diseases such as diabetic retinopathy, psoriasis, restenosis and neovascular glaucoma.

In addition, angiogenesis is also involved in tumor formation and metastasis. For example, tumors larger than about 2 millimeters in diameter have to obtain their own blood source and are known to achieve this by inducing the growth of new capillaries. After these new blood vessels are inserted into the tumor, they provide the nutrients and growth factors essential for tumor growth and promote metastasis of tumor cells.

In order to treat angiogenesis-related diseases, antiangiogenic agents that specifically target angiogenesis have been developed. See, for example, US Pat. No. 6,919,309; US Patent Application Publication 2006/0009412; And PCT Application Publications WO04 / 027027 and WO05 / 117876. In addition, agents that target established vasculature (so-called “Vascular Targeting Agents” or VTAs) have also been developed. Such agents are thought to act by selectively weakening the microtubule cytoskeleton of endothelial cells, thereby severely changing the shape of the cells that occlude blood vessels and disrupt blood flow. See, for example, WO 2005/113532.

Thiocholacin dimers are hydrophobic compounds described previously. See, for example, US Pat. No. 6,627,774. Such compounds have a dual action mechanism, ie antimicrotubule activity and topoisomerase I inhibitory activity (Raspaglio et al., Biochem. Pharmacol . 2005, 69 (1): 113-21). As cytotoxic chemotherapeutic agents for cancer treatment, nanoparticle albumin-binding preparations Nab-5404 and Nab-5676 of thiocolchicine dimers have been developed. For example, Bernnacki et al., Proc . Amer . Assoc . Cancer Res . , vol. 46, 2005 # 2390 and PCT patent application PCT / US2006 / 006167. Intravenous administration of Nab-5404 at 24 mg / kg, qd × 5, has been shown to be able to completely regress tumors and heal in A121 ovarian tumor xenografts.

The disclosures of all documents, patents, patent applications and published patent applications referred to herein are hereby incorporated by reference in their entirety.

Summary of the Invention

The present invention comprises administering to a subject a composition comprising a colchicine or thiocolchicine dimer (eg, IDN5404), the composition being an amount effective to inhibit angiogenesis, wherein the amount of the composition is within the subject Provided are methods for inhibiting angiogenesis in a subject, which are insufficient to cause significant cytotoxicity. In some embodiments, the method comprises administering to a subject a composition comprising an effective amount of a colchicine or thiocolhitine dimer (eg, IDN5404), and wherein the amount (ie, administration) of a colchicine or thiocolhitchin dimer (eg, IDN5404) in the composition Suppression of angiogenesis in a subject having a dose of less than about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 mg / m ² body surface. A method is provided. In some embodiments, the method comprises administering to a subject a composition comprising an effective amount of a colchicine or a thiocolchicine dimer (eg, IDN5404), wherein the amount of the colchicine or thiocolchicine dimer (eg, IDN5404) in the composition is about 0.05, 0.08 A method of inhibiting angiogenesis in a subject is provided that is less than any of 0.1, 0.2, 0.3, 0.4, 0.5 or 0.6 mg / kg. In some embodiments, the composition is about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week or more than one day. Administered. In some embodiments, the composition is administered for any one or more of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more (without interruption or interruption). . In some embodiments, the composition is administered by any of intravenous, intraocular, intraarterial, oral, topical, or inhalation routes.

The composition used in the methods described herein may further comprise a biocompatible polymer such as a carrier protein. For example, in some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the composition is used to inhibit angiogenesis. An effective amount is provided, wherein the amount of the composition is insufficient to cause significant cytotoxicity in the subject. In some embodiments, the method comprises administering to a subject an effective amount of a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), and wherein the colchicine or thiocolhitine dimer (eg, A method of inhibiting angiogenesis in a subject wherein the amount of IDN5404) is less than about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 mg / m ² body surface This is provided. In some embodiments, the method comprises administering to a subject an effective amount of a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), and wherein the colchicine or thiocolhitine dimer (eg, A method of inhibiting angiogenesis in a subject is provided wherein the amount of IDN5404) is less than about 0.05, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5 or 0.6 mg / kg. In some embodiments, the carrier protein containing composition is substantially free of surfactant (eg, free). In some embodiments, the composition is about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week or more than one day. Administered. In some embodiments, the composition is administered for any one or more of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more (without interruption or interruption). . In some embodiments, the composition is administered by intravenous, intraocular, intraarterial, oral, topical, or inhalation route.

Compositions used in the methods described herein can include particles (eg, microparticles or nanoparticles) comprising carrier proteins (eg, albumin) and colchicine or thiocolhitine dimers. For example, in some embodiments, the method comprises administering to a subject a composition comprising a nanoparticle comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), the composition comprising a blood vessel A method effective for inhibiting formation, wherein the amount of the composition is insufficient to cause significant cytotoxicity in the subject, a method of inhibiting angiogenesis in a subject is provided. In some embodiments, the method comprises administering to a subject an effective amount of a composition comprising a nanoparticle comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the colchicine or thiocolhitine in the composition is administered. The amount of dimer (eg, IDN5404) is less than any of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 mg / m ² body surface. A method of inhibiting angiogenesis is provided. In some embodiments, the method comprises administering to a subject an effective amount of a composition comprising a nanoparticle comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the colchicine or thiocolhitine in the composition is administered. A method of inhibiting angiogenesis in a subject is provided wherein the amount of dimer (eg, IDN5404) is less than any of about 0.05, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, or 0.6 mg / kg. In some embodiments, the composition is about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week or more than one day. Administered. In some embodiments, the composition is administered for any one or more of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more (without interruption or interruption). . In some embodiments, the composition is administered by intravenous, intraocular, intraarterial, oral, topical, or inhalation route.

In some embodiments, the average diameter of the particles in the composition is about 200 nm or less. In some embodiments, the particle containing composition is substantially free of surfactant (eg, free). In some embodiments, the weight ratio of protein (eg, albumin) to colchicine or thiocolchicine dimer in the composition is about 18: 1 or less, such as 9: 1 or less. In some embodiments, colchicine or thiocolhitine dimers are coated with a carrier protein (eg, albumin). In some embodiments, the average diameter of the particles in the composition is about 200 nm or less and the composition is substantially free of surfactants (eg, free). In some embodiments, the average diameter of the particles (particularly nanoparticles) in the composition is about 200 nm or less, and colchicine or thiocolchicine dimers are coated with a protein (eg, albumin). Other combinations of the above features are also contemplated. In some embodiments, the particle composition is Nab-5404. Particle compositions comprising other colchicine or thiocolhitine dimers may also include one or more of the above features.

Colchicine or thiocolhitine dimers described herein include two (same or different) subunits of colchicine, thiocolhitine or derivatives thereof. In some embodiments, the colchicine or thiocolchicine dimers comprise one or more thiocolchicine subunits. In some embodiments, the colchicine or thiocolhitine dimer comprises two thiocolhitine subunits. In some embodiments, the colchicine or thiocolchicine dimer is a compound of formula (I)

Wherein B in each subunit is a methoxy or methylthio group, R ₂ is methylenedioxy with methoxy, hydroxyl or R _3, and R ₃ is methylenedi with methoxy, hydroxyl or R ₂ Oxy and X is a linking group. In some embodiments, X contains one or more carbon atoms.

In some embodiments, the colchicine or thiocolchicine dimer is a compound of formula (II)

Wherein B ₁ is a methoxy or methylthio group, B ₂ is a methoxy or methylthio group, n is an integer from 0 to 8, Y is a CH ₂ group, or when n is 1 it is also a formula NH It may be a flag. In some embodiments, n is any one of 0, 1, 2, 3, 4, 5, 6, 7 or 8 (eg, selected from the group consisting of these). In some embodiments n is 1. In some embodiments, n is 1 and Y is NH. In some embodiments n is 2.

In some embodiments, B ₁ and B ₂ are both methoxy groups. In some embodiments, B ₁ and B ₂ are both methylthio groups. In some embodiments, B ₁ is a methoxy group and B ₂ is a methylthio group. In some embodiments, B ₁ is a methylthio group and B ₂ is a methoxy group. In some embodiments, the colchicine or thiocolchicine dimer is any of IDN5404, IDN5676, IDN5800 and IDN5801 (in some embodiments selected from the group consisting of these). In some embodiments, the colchicine or thiocolchicine dimer is IDN5404. In some embodiments, the colchicine or thiocolchicine dimer is IDN5676.

In some embodiments, a composition comprising a nanoparticle comprising albumin and one of IDN5404, IDN5676, IDN5800 and IDN5801 (hereinafter referred to as "Nab-5404,""Nab-5676,""Nab-5800" and "Nab- respectively) A method of inhibiting angiogenesis in a subject is provided, comprising administering to the subject). In some embodiments, the method comprises administering Nab-5404 (or any of Nab-5676, Nab-5800, and Nab-5801) to the subject, and comprising Nab-5404 (or Nab-5676, Nab-5800, and Nab-5801) Of which is an amount effective to inhibit angiogenesis, and the amount of Nab-5404 (or any of Nab-5676, Nab-5800, and Nab-5801) is insufficient to cause significant cytotoxicity in the subject. Phosphorus, methods of inhibiting angiogenesis in a subject are provided. In some embodiments, the method comprises administering to a subject an effective amount of Nab-5404 (or any of Nab-5676, Nab-5800 and Nab-5801), and Nab-5404 (or Nab-5676, Nab-5800 and Nab) Any one of -5801) is less than about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 mg / m ² body surface. A method of inhibiting angiogenesis is provided. In some embodiments, the method comprises administering to a subject an effective amount of Nab-5404 (or any of Nab-5676, Nab-5800 and Nab-5801), and Nab-5404 (or Nab-5676, Nab-5800 and Nab) A method of inhibiting angiogenesis in a subject is provided wherein the amount of any of -5801) is less than any of about 0.05, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5 or 0.6 mg / kg. In some embodiments, the composition is about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week or more than one day. Administered. In some embodiments, the composition is administered for any one or more of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more (without interruption or interruption). . In some embodiments, the composition is administered by intravenous, intraocular, intraarterial, oral, topical, or inhalation route.

The methods described herein are generally useful for the treatment of angiogenesis related diseases. In some embodiments, the angiogenesis related disease may include, for example, ocular disease (eg, macular degeneration, diabetic retinopathy or neovascular glaucoma), cardiovascular disease (eg, restenosis and atherosclerosis), skin disease (eg, psoriasis) ) And arthritis (eg, rheumatoid arthritis).

For example, in some embodiments, comprising administering to a subject a composition comprising a colchicine or a thiocolchicine dimer, wherein the composition is effective in treating a non-tumoral angiogenesis related disease. A method of treating a disease is provided. In some embodiments, the amount of the composition is insufficient to cause significant cytotoxicity in the subject. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), the composition treating the non-neoplastic angiogenesis related disease in the subject. A method effective for treating non-neoplastic angiogenesis related diseases in a subject is provided. In some embodiments, the amount of the composition is insufficient to cause significant cytotoxicity in the subject. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and particles (eg, nanoparticles) comprising a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the composition comprises Methods of treating non-neoplastic angiogenesis related diseases in a subject are provided that are amounts effective to treat non-neoplastic angiogenesis related diseases. In some embodiments, the amount of the composition is insufficient to cause significant cytotoxicity in the subject. In some embodiments, administering Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) to the subject, wherein Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) is splenic Methods of treating non-neoplastic angiogenesis related diseases in a subject are provided that are amounts effective to treat benign angiogenesis related diseases. In some embodiments, the amount of Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) is insufficient to cause significant cytotoxicity in the subject.

In some embodiments, the angiogenesis related disease is a tumor related disease, including, for example, cancer and benign tumors. Cancers that can be treated with the methods described herein include, but are not limited to, breast cancer, colorectal cancer, rectal cancer, non-small cell lung cancer, non-Hodgkin's lymphoma (NHL), renal cell cancer, prostate cancer, liver cancer, pancreatic cancer, soft tissue sarcoma, Kaposi's sarcoma, carcinoid carcinoma, head and neck cancer, melanoma, ovarian cancer, mesothelioma, glioma, glioblastoma, neuroblastoma and multiple myeloma. In some embodiments, the cancer is a solid tumor (eg, metastatic solid tumor). By way of example, methods of treating cancer are further described below.

For example, in some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer, the composition being an amount effective to treat cancer, wherein the amount of the composition is a significant cell in the subject. Methods of treating cancer in a subject are provided that are insufficient to cause toxicity. In some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer, wherein the composition is an amount effective to treat cancer, and the amount of colchicine or thiocolchicine dimer in the composition is colchicine or thiocolhitchin About 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 11%, 12%, 13 of the corresponding maximum tolerated dose of dimer ("MTD") A method of treating cancer in a subject is provided that is less than%, 14% or 15%. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the composition is an amount effective to treat cancer, and A method of treating cancer in a subject is provided wherein the amount of the composition is insufficient to cause significant cytotoxicity in the subject. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the composition is an amount effective to treat cancer, and the composition The amount of colchicine or thiocolhitine dimer in the sample is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 11% of the corresponding MTD of the colchicine or thiocolchicine dimer. A method of treating cancer in a subject is provided that is less than 12%, 13%, 14% or 15%. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and particles (eg, nanoparticles) comprising a colchicine or thiocolhitine dimer (eg, IDN5404), wherein the composition comprises A method effective for treating cancer, wherein the amount of the composition is insufficient to cause significant cytotoxicity in the subject. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and particles (eg, nanoparticles) comprising a colchicine or thiocolhitine dimer (eg, IDN5404), and colchicine in the composition Or the amount of thiocolchicine dimer is about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 11%, 12 of the colchicine or the corresponding MTD of the thiocolchicine dimer. A method of treating cancer in a subject is provided that is less than%, 13%, 14% or 15%. In some embodiments, administering Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) to a subject, wherein Nab-5404 (or Nab-5676, Nab-5800 or Nab-58Ol) is cancerous A method effective for treating cancer in a subject, wherein the amount of Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) is insufficient to cause significant cytotoxicity in the subject. do. In some embodiments, the method comprises administering Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) to the subject, and Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) is cancerous. Is an effective amount to treat the amount of Nab-5404 (or Nab-5676, Nab-5800 or Nab-5801) to the corresponding MTD of Nab-5404 (or Nab-5676, Nab-5800, or Nab-5801). Cancer in an individual, less than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 11%, 12%, 13%, 14%, or 15% The treatment method of is provided.

Also provided are pharmaceutical compositions, unit dosages, kits, and products useful in the methods described herein.

It will be appreciated that one, some or all of the features of the various embodiments described herein may be combined to form other embodiments of the invention.

Brief description of the drawings

1A shows the effect of Nab-5404 on microtubule formation using MX-1 breast carcinoma cells. 1B shows the effect of Nab-5676 on microtubule formation using MX-1 breast carcinoma cells.

2A-2C are micrographs of stained microtubule networks in MX-1 cells after treatment with control vehicle (PBS, 2A), 0.6 μg / ml Nab-5404 (2B) and μg / ml Nab-5676 (2C) Indicates.

3 shows antiangiogenic activity of Nab-5404, Nab-5676 and CA4P as assessed by microvascular / tubule formation. Cells were incubated with various compounds on day 1 and stained on day 12. 3A-3D show tubule formation micrographs after treatment with control vehicle (3C), 0.01 μg / ml Nab-5404 (3A), 0.01 μg / ml Nab-5676 (3B) and 0.01 μg / ml CA4P (3D). to provide.

4 shows a comparison of tubule length for cells treated with Nab-5404, Nab-5676 and CA4-P over a concentration range of 0.0-100 μg / ml.

5 shows antiangiogenic activity of Nab-5404 and Nab-5676 as assessed by microvascular / tubule formation or destruction. Cells were incubated with various compounds on day 8 and stained on day 11. 5A-5C provide micrographs of tubule formation after treatment with control vehicle (5A), 0.01 μg / ml Nab-5404 (5B) and 0.01 μg / ml Nab-5676 (5C). 4D shows a comparison of tubule length for cells treated with Nab-5404 and Nab-5676 over a concentration range of 0.0-10 μg / ml.

6 shows antiangiogenic activity of Nab-5404 as assessed by microvascular / tubule formation or destruction. Cells were incubated with various concentrations of compound on day 11 and stained on day 12. 6A-6F show control vehicle (6A), 0.001 μg / ml Nab-5404 (6B), 0.01 μg / ml Nab-5404 (6C), 0.1 μg / ml Nab-5404 (6D), 1 μg / ml Nab- Micrographs of tubule formation and destruction after treatment with 5404 (6E) and 10.0 μg / ml Nab-5404 (6F) are provided.

7 shows the antiangiogenic activity of Nab-5676 as assessed by microvascular / tubule formation or destruction. Cells were incubated with various concentrations of compound on day 11 and stained on day 12. 7A-7F show control vehicle (7A), 0.001 μg / ml Nab-5676 (7B), 0.01 μg / ml Nab-5676 (7C), 0.1 μg / ml Nab-5676 (7D), 1 μg / ml Nab- Micrographs of tubule formation and destruction are shown after treatment with 5676 (7E) and 10.0 μg / ml Nab-5676 (7F).

8 shows antiangiogenic activity of CA4P assessed by microvascular / tubule formation or destruction. Cells were incubated with various concentrations of compound on day 11 and stained on day 12. 8A-8C show micrographs of tubule formation and destruction after treatment with 0.01, 0.1, and 1.0 μg / ml CA4P, respectively.

9 shows a comparison of the antiangiogenic activity of Nab-5404, Nab-5676 and CA4P assessed by microvascular or tubule formation or destruction. Cells were incubated with compounds on day 11 and stained on day 12. This figure shows the tubule length for cells treated with each compound over a concentration range of 0.0 to 10 μg / ml.

10 shows the effect of Nab-5404 on HT-29 tumor growth in a xenograft murine model using two cycles of low dose / high dose schedule. The first cycle (0-14 days) administration was 1.7, 2.5 or 3.4 mg / kg and the second cycle (15-30 days) administration was 20, 30 or 40 mg / kg. 10A shows the mean tumor volume (n = 10) from day 0 to day 40. 1OB shows the mean weight loss percentage of mice over 0-40 days.

FIG. 11 shows the effect of Nab-5676 on HT-29 tumor growth in a xenograft murine model using two cycles of low dose / high dose schedule. The first cycle (0-14 days) administration was 1.7, 2.5 or 3.4 mg / kg and the second cycle (15-30 days) administration was 20, 30 or 40 mg / kg. 11A shows the mean tumor volume (n = 10) from day 0 to day 40. 11B shows the mean weight loss percentage of mice over 0-40 days.

12 shows the effect of CA4P on HT-29 tumor growth in xenograft murine models. HT-29 tumors were treated at 100 mg / kg after being 900 mm ³ in volume. 12A shows mean tumor volume (n = 10) from day 28 to day 41 (square = vehicle control, diamond = CA4P treatment). 12B shows the mean weight loss percentage of mice over 28-41 days (square = vehicle control, diamond = CA4P treatment).

Detailed description of the invention

The invention thio colchicine composition comprising the dimer, in particular, thio colchicine albumin-containing nanoparticle preparation of the dimer, and more specifically, in vitro (in vitro) IDN-5404 nanoparticle albumin-containing formulations ( "Nab- in 5404 ") and albumin containing nanoparticle formulations of IDN-5676 (" Nab-5676 ") are effective in inhibiting microvascular formation and are based in part on the observations of the inventors. Triu et al., 97th AACR Annual Meeting, Abstract No. 3823]. IC50 values for these activities are significantly lower than those required for in vitro cytotoxic activity of the composition. The inventors further note that compositions comprising thiocolchicine dimers, in particular, albumin containing nanoparticle formulations of thiocolchicine dimers (e.g., Nab-5404 and Nab-5676), have a corresponding maximum tolerable dose (MTD) of the composition. in vivo (in a much lower dose than in vivo ), it was observed to be effective in preventing tumor growth. This observation means that a composition comprising thiocolchicine dimer or an analog thereof (eg, colchicine dimer) has antiangiogenic and vascular targeting activity independent of its cytotoxic effect. When administered in an effective and non-cytotoxic amount, the composition can optionally target the growth of new blood vessels and block blood flow without causing significant cell death in the target tissue.

Effective and non-cytotoxic amounts of colchicine or thiocolchicine dimers (ie, insufficient to cause significant cytotoxicity) are desirable for targeting non-tumor angiogenesis related diseases in order to minimize the cytotoxic effects of the compounds. Effective and noncytotoxic amounts are also advantageous for treating cancer. Traditional chemotherapy with cytotoxic agents is typically carried out in an amount equal to or similar to the maximum allowable dose of the agent to minimize the cytotoxic effects of the agent. However, this high dose schedule requires a long period of untreatment to restore normal host cells. On the other hand, tumor cells can also resume growth during the untreated period. This may increase the risk of drug resistant tumor cell development. Non-cytotoxic doses (ie amounts) of colchicine or thiocolchicine dimers reduce the risk of developing drug resistance because treatment does not significantly stop during treatment cycles. In addition, non-cytotoxic dosages minimize the side effects caused by the drug and the likelihood of developing apparent systemic toxicity (eg, weight loss).

Thus, the present invention comprises administering to a subject a composition comprising a colchicine or thiocolchicine dimer in one aspect, the composition being an amount effective to inhibit angiogenesis, wherein the amount of the composition is a significant cell in the subject. Provided are methods for inhibiting angiogenesis in a subject, including established angiogenesis targeting, that are insufficient to cause toxicity. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein (eg, albumin) and a colchicine or thiocolhitine dimer, wherein the composition is an amount effective to inhibit angiogenesis, the composition The amount of is insufficient to cause significant cytotoxicity in the subject.

In another aspect, the invention comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer, wherein the composition is an amount effective for treating a non-tumour angiogenesis related disease. Provides a method of treatment. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein and a colchicine or thiocolchicine dimer, wherein the composition is an amount effective for treating a non-neoplastic angiogenesis related disease.

In another aspect, the invention comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer, wherein the composition is an amount effective for treating a tumor associated disease, the amount of the composition being a significant cell in the subject. Provided are methods of treating tumor associated diseases in a subject that are insufficient to cause toxicity. In some embodiments, the method comprises administering to a subject a composition comprising a carrier protein and a colchicine or thiocolhitine dimer, wherein the composition is an amount effective to treat cancer, wherein the amount of the composition is significant within the subject. Insufficient to cause cytotoxicity.

Also provided are pharmaceutical compositions, unit dosages, kits, and products useful in the methods described herein.

In general, "composition" or "compositions" comprise compositions of the invention and may be applied to compositions of the invention. The invention also provides a pharmaceutical composition comprising the components described herein.

The term “individual” is a mammal, including humans. Individuals include, but are not limited to, humans, cattle, horses, cats, dogs, rodents, or primates. In some embodiments, the subject is a human. In some embodiments, the subject is an experimental animal model for studying angiogenesis or angiogenesis related disease (s).

The present invention includes all stereoisomers, including enantiomers and diastereomers of the compounds mentioned herein. Unless stereochemistry is explicitly indicated in structure, the disclosed structure is to encompass all possible stereochemical variations. The present invention encompasses all enantiomers, substantially pure situs or preferential forms of any of the chiral compounds disclosed, or racemic mixtures, or any proportion of enantiomers. The present invention includes any diastereomeric, substantially pure diastereomeric forms and mixtures of all proportions of the compounds mentioned in the above formula. The invention also includes all solvates of the compounds mentioned herein, including all hydrates of the compounds mentioned herein. The invention also includes all polymorphs, including the crystalline and amorphous forms of the compounds mentioned herein. The invention also includes all salts of the compounds mentioned herein, in particular pharmaceutically acceptable salts. Metabolites and prodrugs of the compounds disclosed herein are also included in the present invention. For all uses of the compounds disclosed herein, the invention also relates to stereochemistry, enantiomers, diastereomers, solvates, hydrates, polymorphs, crystals, amorphous, salts, pharmaceutically acceptable salts of the compounds as described. , Any or all use of metabolites and prodrug variants.

Aspects and embodiments of the invention described herein are to be understood to include and consist of and / or consist essentially of aspects and embodiments.

Inhibition of angiogenesis

The present invention provides, in one aspect, a method of inhibiting angiogenesis in a subject by administering a composition comprising a colchicine or thiocolchicine dimer (eg, a carrier protein containing composition). The composition is an amount effective to inhibit angiogenesis. However, the amount of the composition administered is insufficient to cause significant cytotoxicity in the subject.

As used herein, “angiogenesis” refers to the process of forming new blood vessels in a tissue or organ. Angiogenesis typically begins by erosion of the vascular basement membrane by enzymes released by endothelial cells and leukocytes. Thereafter, endothelial cells lining the lumen of the blood vessel protrude through the basement membrane. Angiogenic stimuli cause endothelial cells to migrate through the eroded basement membrane. Migrating cells form "sprouts" of the vascular blood vessels, where mitosis and proliferation of endothelial cells occur. Endothelial processes merge together to form capillary loops and create new blood vessels.

"Inhibition of angiogenesis" means reducing, obstructing or inhibiting angiogenesis in one or more tissues in an individual, including, for example, ocular tissue, cardiovascular tissue, skin tissue, arthritic tissue and tumor tissue. Inhibition of angiogenesis affects one or more stages of the angiogenesis process, for example, attenuates the migration and survival of activated endothelial cells, prevents “protrusion” of cells from the maternal vessels, and / or forms new blood vessels It can be achieved through the prevention of. Changing the density of microvessels is also included in the term “inhibiting angiogenesis”. The term “inhibition of angiogenesis” also includes destroying established vasculature. "Destroys an established vasculature" means the ability to occlude, dissolve or otherwise affect the vasculature formed and formed by angiogenesis. The destruction of the vascular system can be reversible or irreversible, and can be partial or complete.

Thus, the methods provided herein include one or more of the following aspects: inhibition (eg, reduction, interference or prevention) of endothelial cell migration, inhibition (eg, reduction, interference or prevention) of endothelial cell "protuberances" from the mother blood vessels. ), Inhibition (eg, reduction, obstruction or prevention) of new angiogenesis and targeting of an established vasculature formed by angiogenesis (eg, occlusion, destruction or collapse). In some embodiments, methods of inhibiting (eg, reducing, obstructing or preventing) blood vessel formation in a subject are provided. In some embodiments, methods of disrupting (eg, occluding, dissolving, or otherwise affecting) an established vascular system are provided. In some embodiments, a method of reducing the density of microvessels in a tissue of an individual is provided.

An "effective amount" refers to an amount or dosage of a composition that is administered to a subject in a single or multiple doses to provide the desired effect in the subject. For example, if the amount of the composition is sufficient to reduce, interfere with, or prevent angiogenesis (eg, one or more aspects of angiogenesis) in one or more tissues in a subject, the composition is an “effective amount to inhibit angiogenesis”. As used in the context of treatment, the term “effective amount” refers to an amount sufficient to treat a specified disorder, condition or disease, such as an amount of a compound or composition sufficient to ameliorate, alleviate, alleviate and / or delay one or more symptoms. Effective amounts can be determined in vitro and / or in vivo. Methods for determining the effective amount of a composition for inhibiting angiogenesis are known in the art.

In some embodiments, the amount of the composition is effective for inhibiting angiogenesis in eye tissue. In some embodiments, the amount of the composition is effective for inhibiting angiogenesis in cardiovascular tissue. In some embodiments, the amount of the composition is effective for inhibiting angiogenesis in skin tissue. In some embodiments, the amount of the composition is effective for inhibiting angiogenesis in arthritic tissue. In some embodiments, the amount of the composition is effective for inhibiting angiogenesis in tumor tissue. In some embodiments, the amount of the composition is effective for inhibiting new blood vessel formation. In some embodiments, the amount of the composition is effective for inhibiting "protuberance" of endothelial cells from the parent blood vessel. In some embodiments, the amount of the composition is effective for inhibiting endothelial cell migration. In some embodiments, the amount of the composition is effective to destroy an established vasculature. In some embodiments, the amount of the composition is effective to inhibit angiogenesis (eg, any one or more aspects of angiogenesis) to at least about 5%, 10%, 20%, 40%, 50% or more.

If the amount of the composition is an amount sufficient for colchicine or thiocolhitine dimer to cause significant cell death in the individual, the composition is referred to as "in insufficient amount to cause significant cytotoxicity" (also referred to as "non-cytotoxic amount"). ). Cytotoxicity can be measured by one or more of the following. For example, noncytotoxic amounts can be determined based on in vitro cell viability assays. The non-cytotoxic amount may be insufficient to cause at least about 50% cell death in an in vitro cell viability assay. In some embodiments, the amount of the composition is at least about 40%, at least 30%, at least 20%, at least 10%, at least 5%, at least 4%, at least 3%, at least 2%, or 1% in an in vitro cell viability assay. It may be insufficient to cause any of the above cell deaths. In some embodiments, the amount of the composition may be insufficient to cause any measurable cell death in an in vitro cell viability assay. Suitable cells for in vitro viability assays include, but are not limited to, tumor cells (eg, MX-1 breast carcinoma cell line, HepG2 hepatocarcinoma cell line, and HT-29 colon carcinoma cell line) and normal cells (eg primary rat hepatocytes). do. The noncytotoxic amount can also be determined based on an in vivo analysis of drug toxicity. For example, the noncytotoxic amount may be an amount sufficient to kill at least about 50% of the test population in an in vivo cytotoxicity assay. In some embodiments, the amount of colchicine or thiocolchicine dimer is at least about 40%, at least 30%, at least 20%, at least 10%, at least 5%, at least 4%, at least 3% of the test population in an in vivo cytotoxicity assay. Insufficient to kill either at least 2% or at least 1%. In some embodiments, the amount of the composition is insufficient to cause any killing of the test population in an in vivo drug toxicity assay. The non-cytotoxic amount can also be determined based on the amount of colchicine or thiocolchicine dimer required to cause apparent systemic toxicity (eg, weight loss) in the subject, i.e., if the drug does not cause any apparent systemic toxicity The amount of is noncytotoxic. For example, in some embodiments, the non-cytotoxic amount is an amount that causes a weight loss of less than about 15% (including, for example, less than about 10%, 8%, 5% or less).

In some embodiments, the amount of colchicine or thiocolchicine dimer in the composition at each administration is about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 and 15 mg / m ² Which is less than. For example, the amount of colchicine or thiocolchicine dimer can range from about 0.25 mg / m ² body surface to about 15 mg / m ² , about 0.25 mg / m ² to about 10 mg / m ² , about 0.25 mg / m ² to about 8 mg / m ² , about 0.25 mg / m ² to about 4 mg / m ² and about 0.25 mg / m ² to about 2 mg / m ² . In some embodiments, the amount of colchicine or thiocolchicine dimer in the composition at each administration is less than about 0.05, 0.08, 0.1, 0.2, 0.3, 0.4, 0.5, or 0.6 mg / kg. For example, the amount of colchicine or thiocolchicine dimer may range from about 0.05 mg / kg to about 0.5 mg / kg, about 0.08 mg / kg to about 0.3 mg / kg, and about 0.1 mg / kg to about 0.2 mg / mg Can be.

In some embodiments, colchicine and thiocolhitine dimers are amounts that do not develop drug resistance for long periods of time (eg, 6 months or longer) that are stopped or uninterrupted. In some embodiments, colchicine and thiocolchicine dimers are amounts that do not cause significant side effects (eg, side effects typically associated with chemotherapy) in an individual. Side effects typically associated with chemotherapy include, for example, dehydration, diarrhea, nausea, vomiting, loss of vision or disturbances, and anemia.

The frequency of administration of the composition is not limited thereto, but at least about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week or once daily Which frequency is included. In some embodiments, the interval between each administration is less than about 1 week, such as less than 6, 5, 4, 3, 2 or 1 day. In some embodiments, the interval between each administration is constant. For example, the administration can be performed daily, every 2 days, every 3 days, every 4 days, every 5 days, or weekly. In some embodiments, administration can be performed twice daily, three times daily, or more often.

Administration of the composition can be extended over a long period of time, such as from about 1 month to about 3 years. For example, the dosing regimen can be extended over any of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30 and 36 months. In some embodiments, there is no interruption in the dosing schedule. In some embodiments, the interval between each administration is about 1 week or less.

The compositions described herein include, but are not limited to, intravenous, intraperitoneal, intraocular, intraarterial, pulmonary, oral, alveolar, intramuscular, intratracheal, subcutaneous, intraocular, intratracheal, transdermal, pleural transit, intraarterial, Topical, inhalation (eg, as a mist of spray), intramucosal (eg, via the nasal mucosa), subcutaneous, transdermal, gastrointestinal, intra-articular, intra-tank, intraventricular, rectal (ie, via suppositories), vagina It may be administered to the subject via any route of the art, including intracranial, intraurethral, intrahepatic, and intratumoral (ie via vaginal suppositories). In some embodiments, the composition is toured systemically. In some embodiments, the composition is administered topically. In some embodiments, the composition is administered by intravenous, intraocular, intraarterial, oral, topical, or inhalation route.

If the method relates to the inhibition of angiogenesis in eye tissue, the composition may be administered directly to the eye or eye tissue. The composition may be administered topically to the eye as eye drops. The composition may also be administered by injection into the eye or tissues associated with the eye. The composition may be intraocular injection, perioperative injection, subretinal injection, intravitreal injection, transseptal injection, subscleral injection, intrachoroidal injection, intra anterior injection, subconjunctival injection, subtennocentral injection, post-ocular injection, eyelid injection, or Administration via posterior scleral delivery. The composition can be administered, for example, to the glassy, waterproof, sclera, conjunctiva, the site between the sclera and the conjunctiva, the retinal choroid tissue, the macula, or other site within or adjacent to the subject's eye. For an exemplary description of the perioperative route for retinal drug delivery, see Periocular routes for retianl drug delivery, Raghava et al. (2004), Expert Opin . Drug Deliv , 1 (1): 99-114. The composition may also be administered to the subject as an implant. Preferred implants are biocompatible and / or biodegradable sustained release formulations that release the compound slowly over long periods of time. Eye implants for drug delivery are known in the art. See, for example, U.S. Patent Applications 5,501,856, 5,476,511 and 6,331,313. The compositions may also be administered to a subject using iontophoresis, including but not limited to, iontophoresis described in US Pat. Nos. 4,454,151 and US Patent Application Publication Nos. 2003/0181531 and 2004/0058313.

Methods of treating angiogenesis related diseases

The methods described herein are generally useful for the treatment of angiogenesis related diseases. "Angiogenesis related disease" refers to a disease or disorder in which angiogenesis is an aspect of the disease. Angiogenesis-related diseases may be caused by abnormal angiogenesis. In some embodiments, the angiogenesis related disease is mediated at least in part by angiogenesis. In some embodiments, angiogenesis is essential for the development of angiogenesis related diseases. Angiogenesis related diseases are known in the art and include, for example, non-tumoral angiogenesis related diseases such as macular degeneration, diabetic retinopathy, rheumatoid arthritis, and other diseases described herein. In some embodiments, the angiogenesis related disease is a tumor related disease such as cancer or a benign tumor.

As used herein, “treatment” is a method for obtaining beneficial or desired clinical results. For the purposes of the present invention, beneficial or desired clinical outcomes include the relief of one or more symptoms, a reduction in the spread of the disease, a stabilized (ie, not worsening) condition of the disease, prevention or delay of disease spread, prevention of disease occurrence or recurrence. Or delay, slow or slow the progression of the disease, ameliorate the disease state, and alleviate (either partially or in total). "Treatment" also includes reducing the pathological prognosis of angiogenesis related diseases. The methods of the present invention contemplate any one or more of the above aspects of treatment.

Non-tumour  Treatment of Angiogenesis-Related Diseases

The methods described herein are useful for the treatment of non-tumoral angiogenesis related diseases.

In some embodiments, the method is useful for the treatment of non-tumoral angiogenesis related diseases in eye tissues such as cornea, retina, macula and choroid. Such methods are generally useful for preventing blindness, vision loss (eg, loss of visual intelligibility or vision) and / or other prognosis in various ocular diseases. In some embodiments, methods of treating macular degeneration, including senile macular degeneration (AMD), are provided. AMD is characterized by progressive loss of central vision, caused by damage to photoreceptor cells in the retina, called the macula. AMD is largely divided into two clinical states, wet and dry, with dry forms accounting for 80-90% of all cases. The dry form is characterized by superficial atrophy characterized by RPE cell death, with photoreceptor atrophy clinically present and with superficially deposited macular microsphere disease between the retinal pigment epithelium (RPE) and the Bruch's membrane. Wet AMD, which causes approximately 90% of severe vision loss, is associated with angiogenesis in the macular area and leakage of these new blood vessels. Accumulation of blood and fluid can cause the retina to detach, resulting in rapid photoreceptor degeneration and vision loss. It is generally known that the dry form of AMD precedes the wet form.

The methods provided herein are particularly useful for treating or inhibiting wet forms of macular degeneration. Inhibition of angiogenesis in the ocular tissue also prevents or delays the transition of the macular degeneration to the wet form of the macular degeneration. The present invention therefore also provides a method of treating the dry form of macular degeneration. The invention also includes, but is not limited to, methods of treating or preventing one or more aspects or symptoms of macular degeneration, including photoreceptor cell loss, loss of vision (including, for example, visual clarity and visual field) and retinal detachment. Other related aspects such as photoreceptor degeneration, RPE degeneration, retinal degeneration, choroidal degeneration, cone degeneration, retinal dysfunction, retinal damage, Bruch's membrane damage, loss of RPE function, extracellular matrix of normal macular and / or tissue structure of cells Loss of integrity, loss of function of cells in the macular and photoreceptor dystrophy are also included.

Other non-neoplastic angiogenesis related ocular diseases that can be treated by the methods described herein include, but are not limited to, angiogenesis of the retina, angiogenesis of the cornea (eg, trachoma, infection, inflammation, transplantation or trauma caused by trauma). Diabetic retinopathy, diabetic retinopathy, diabetic macular edema, ischemic retinopathy, hypertensive retinopathy, obstructive retinopathy, immature retinopathy, post-traumatic angiogenesis, angiogenesis after infection, angiogenesis after transplantation, retinal detachment or retina Angiogenesis after degeneration, neovascular glaucoma, anterior and / or anterior angle angiogenesis, choroidal neovascularization (CNV), subretinal angiogenesis, posterior capsular fibrosis, ocular histoplasmosis syndrome, myopia degeneration, angioplasty, uveitis, Skin flushing, posterior capsular fibrosis, ocular histoplasmosis, and idiopathic central serous chorioretinopathy. In some embodiments, the ocular disease is diabetic retinopathy. In some embodiments, the ocular disease is neovascular glaucoma.

In some embodiments, methods of treating cardiovascular diseases such as atherosclerosis, restenosis, atherosclerosis and hemangiomas are provided. Atherosclerosis is a form of chronic vascular injury and changes in the characteristics of some normal vascular smooth cells (VSMC) in the arterial wall resulting in a dense network of capillaries in atherosclerotic plaques. These fragile microvessels can cause bleeding, produce blood clots, reduce blood flow to the myocardium, and cause heart attacks. Restenosis typically occurs after coronary artery bypass surgery, endoscopic resection and heart transplantation, particularly after cardiac instrument angioplasty, resection, laser ablation, or intravascular stenting. This entails extensive growth of microvessels. By inhibiting angiogenesis in cardiovascular tissues, the methods provided herein are useful for treating such cardiovascular diseases.

In some embodiments, the method is useful for the treatment of rheumatoid arthritis. Rheumatoid arthritis causes the blood vessels in the joints to form angiogenesis, forming extensively distributed blood vessels that penetrate and destroy cartilage. By inhibiting angiogenesis in arthritic tissues, the methods provided herein are useful for treating rheumatoid arthritis. Also provided are methods of treating hemophilia factor joints.

In some embodiments, the method is an angiogenesis-related skin disease, including but not limited to psoriasis, scleroderma, angiogenesis as a prognosis for infections (eg, cat scratches, bacterial ulcers, etc.) and other skin disorders. Useful in the treatment of In some embodiments, the method is useful for the treatment of psoriasis. Psoriasis is a chronic skin disease that occurs in approximately 3% of the world's population. Histological studies, including electron microscopy, have shown that psoriasis is a prominent feature of changes in skin angiogenesis. Therefore, inhibition of angiogenesis in skin tissue is useful for the treatment of psoriasis.

Other angiogenesis related diseases that can be treated with the methods of the present invention include, but are not limited to, Osler-Webber syndrome, hereditary hemorrhagic vasodilation, plaque angiogenesis, capillary dilatation, angiofibromas, hemangiomas, Wound granulation, endometriosis and the like. The present invention is also useful for the treatment of nasal polyposis, especially in cystic fibrosis patients.

In addition to treating angiogenesis related diseases, the methods described herein can also be used to modulate or prevent the occurrence of normal physiological conditions associated with angiogenesis. For example, the methods of the present invention can be used to attenuate angiogenesis associated with ovulation, embryo implantation, placental formation, and the like, and are therefore useful for pregnancy control purposes.

Provided herein are methods of treating the various diseases described herein. In some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer (eg, a composition comprising a carrier protein), wherein the composition is an amount effective to treat a non-tumor angiogenesis related disease. Methods of treating non-neoplastic angiogenesis related diseases in a subject are provided. In some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer (eg, a composition comprising a carrier protein), wherein the composition is an amount effective for treating an ocular disease. Methods of treating related eye diseases (including, for example, macular degeneration, diabetic retinopathy or neovascular glaucoma) are provided. In some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer (eg, a composition comprising a carrier protein), wherein the composition is an amount effective for treating a cardiovascular disease. (Eg, restenosis and atherosclerosis) are provided. In some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer (eg, a composition comprising a carrier protein), wherein the composition is an amount effective for treating a skin condition. Provided are methods of treating a disease (eg, psoriasis). In some embodiments, the method comprises administering to a subject a composition comprising a colchicine or a thiocolchicine dimer (eg, a composition comprising a carrier protein), wherein the composition is an amount effective to treat arthritis (eg, rheumatoid) (Arthritis) is provided.

In some embodiments, the colchicine or thiocolchicine dimer composition is about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week, or It is administered more than any day of the day. In some embodiments, the interval between each administration is less than about 1 week, such as less than about 6, 5, 4, 3, 2, or 1 day. In some embodiments, the interval between each administration is constant. For example, the administration can be performed daily, every 2 days, every 3 days, every 4 days, every 5 days, or weekly. In some embodiments, administration can be performed twice daily, three times daily or more often. Administration of the composition can be extended over an extended period of time (without interruption or interruption), such as over about 1 month to about 3 years. For example, the dosing regimen (without interruption or interruption) can be administered over any of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, and 36 months. Can be extended. In some embodiments, there is no interruption in the dosing schedule. In some embodiments, the interval between each administration is about 1 week or less.

Method of treating tumor-related diseases by inhibition of angiogenesis

The methods described herein are also useful for inhibiting angiogenesis in tumor tissues and treating tumor related diseases such as cancer and benign tumors.

Inhibition of angiogenesis (eg, by inhibiting new angiogenesis or by targeting established blood vessels) prevents the tumor from being supplied with sufficient nutrients and oxygen to aid growth beyond a certain size. Since angiogenesis involves primary tumor growth and metastasis, the methods provided herein can inhibit neoplastic growth of the tumor at the primary site and prevent tumor metastasis at the secondary site. In some embodiments, colchicine or thiocolhitine dimers inhibit new blood vessel formation. In some embodiments, the colchicine or thiocolchicine dimer composition destroys an established tumor vasculature. For example, the composition may be effective for selective occlusion, dissolution of the tumor vasculature, or may otherwise be reversible or irreversible, partially or completely affecting (sometimes proliferating the tumor vasculature).

Tumor-related diseases through the administration of an effective amount of a composition comprising colchicine or thiocolhitine dimers (the amount of the composition is insufficient to cause significant cytotoxicity (also referred to as a "non-cytotoxic dose")) Provides a method of treatment. Traditional chemotherapy with cytotoxic agents is typically carried out at a dose equal to or similar to the maximum allowable dose of the agent to maximize the cytotoxic effect of the agent. However, this high dose schedule requires a long period of untreatment to recover normal host cells. On the other hand, tumor cells can also resume growth during the untreated period. This may increase the risk of drug resistant tumor cell development. Non-cytotoxic doses of colchicine or thiocolchicine dimers reduce the risk of developing drug resistance, since the treatment of cancer does not significantly stop treatment during the treatment cycle.

To provide an example, the methods of treating cancer are further described herein. Such a description is generally understood to apply to all tumor related diseases, including benign tumors.

In some embodiments, the method comprises administering to a subject a composition comprising colchicine or a thiocolchicine dimer, wherein the composition is an amount effective to treat cancer, and the amount of colchicine or thiocolchicine dimer in the composition per administration is the same or About 1%, 2%, 3%, 4%, 5%, 6% of the MTD corresponding to the same (or similar) colchicine or thiocolchicine dimer in the same (or similar) formulation following a similar dosing schedule or a traditional dosing schedule A method of treating cancer in a subject is provided that is less than any of 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% or 15%. In some embodiments, the amount of colchicine or thiocolchicine dimer in the composition per administration is, for example, about 1% to about 12%, about 1% to about 10%, about 1% to about 8%, about 1 of the corresponding MTD. About 1% to about 15% of the corresponding MTD, including any of% to about 5%, about 1% to about 3%. MTDs for colchicine or thiocolchin dimers are known or can be readily determined by one skilled in the art. For example, the MTD for Nab-5404 following the weekly schedule is about 90 to 100 mg / m ² body surface.

In some embodiments, the colchicine or thiocolchicine dimer composition is about once every three weeks, once every two weeks, once a week, twice a week, three times a week, four times a week, five times a week, six times a week, or It is administered more than any day of the day. In some embodiments, the interval between each administration is less than about 1 week, such as less than about 6, 5, 4, 3, 2, or 1 day. In some embodiments, the interval between each administration is constant. For example, the administration can be performed daily, every 2 days, every 3 days, every 4 days, every 5 days, or weekly. In some embodiments, administration can be performed twice daily, three times daily or more often. Administration of the composition can be extended over an extended period of time (without interruption or interruption), such as over about 1 month to about 3 years. For example, the dosing regimen (without interruption or interruption) can be administered over any of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, and 36 months. Can be extended. In some embodiments, there is no interruption in the dosing schedule. In some embodiments, the interval between each administration is about 1 week or less.

Cancers that can be treated with the methods of the present invention include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More specific examples of such cancers include, but are not limited to, squamous cell cancer, lung cancer (including small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma and lung squamous cell carcinoma), peritoneal cancer, hepatocellular cancer, stomach or gastrointestinal cancer (Including gastrointestinal cancer), pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, liver cancer, breast cancer, colon cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney or kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, Liver carcinoma, head and neck cancer, B-cell lymphoma (low grade / follicular non-Hodgkin's lymphoma (NHL), small lymphocyte (SL) NHL, medium grade / follicular NHL, medium grade NHL, high grade immunoblast NHL, high grade) Lymphocytic NHL, High Grade Bovine Non-Cutting Cell NHL, Giant Disease NHL, Mantle Cell Lymphoma, AIDS-Related Lymphoma and Waldenstrom Macroglobulinemia), Chronic Lymphocytic Leukemia (CLL), Acute Lymphocytic Leukemia (ALL), Hair Cells leukemia , Chronic myelogenous leukemia, and post-transplant lymphoproliferative disorders (PTLD), as well as abnormal vascular proliferation, edema (eg, those associated with brain tumors), and MAG syndromes associated with nevus. The methods described herein are particularly useful for the treatment of cancers involving a wide range of angiogenesis, such as squamous cell carcinoma.

In some embodiments, methods of treating primary tumors are provided. In some embodiments, a method of treating metastatic cancer (ie, cancer that has metastasized from a primary tumor) is provided. In some embodiments, a method of treating cancer at an advanced stage is provided. In some embodiments, methods of treating breast cancer (which may be HER2-positive or HER2-negative) are provided, including, for example, advanced breast cancer, stage IV breast cancer, locally advanced breast cancer, and metastatic breast cancer. In some embodiments, the cancer is lung cancer, including, for example, non-small cell lung cancer (NSCLC, such as advanced NSCLC), small cell lung cancer (SCLC, such as advanced SCLC), and advanced solid malignancy in the lung. In some embodiments, the cancer is ovarian cancer, head and neck cancer, gastric malignancy, melanoma (including metastatic melanoma), colorectal cancer, pancreatic cancer, and solid tumor (eg, progressive solid tumor). In some embodiments, the cancer is breast cancer, colorectal cancer, rectal cancer, non-small cell lung cancer, non-Hodgkin's lymphoma (NHL), renal cell cancer, prostate cancer, liver cancer, pancreatic cancer, soft tissue sarcoma, Kaposi's sarcoma, carcinoid carcinoma, head and neck cancer, Melanoma, ovarian cancer, mesothelioma, glioma, glioblastoma, neuroblastoma, and multiple myeloma (in some embodiments selected from the group consisting of these). In some embodiments, the cancer is a solid tumor.

The method described herein can be carried out in an adjuvant setting. "Assistant settings" refer to clinical settings in individuals who have a history of cancer and who have generally responded to therapies, including (but not necessarily) surgery (eg, surgical resection), radiotherapy and chemotherapy, and the like. However, because of the history of cancer, these individuals are considered at risk of developing the disease. Treatment or administration in a "assistant setting" refers to a subsequent mode of treatment. The degree of risk (ie, when an individual in an adjuvant setting is considered "high risk" or "low risk") depends on a number of factors, most commonly the degree of disease when first treated. The methods provided herein can also be practiced with neoadjuvant settings, ie, the methods can be performed before primary / limiting therapy. In some embodiments, the subject has previously been treated. In some embodiments, the individual has not been previously treated. In some embodiments, the treatment is primary therapy.

The methods described herein can be performed in association with other cancer treatment methods (eg, in combination with therapy settings). For example, in some embodiments, the method comprises first line therapy, and second line therapy of administering a composition comprising a colchicine or thiocolchicine dimer. In some embodiments, the second therapy is chemotherapy. In some embodiments, the secondary therapy is radiation therapy. In some embodiments, the second therapy is surgery. Primary and secondary therapies may be performed simultaneously or sequentially (ie, the primary therapy is performed before or after the secondary therapy). In some embodiments, the method comprises co-administering a composition comprising a colchicine or thiocolchicine dimer with a secondary chemotherapeutic agent (simultaneously or sequentially) to the subject, wherein the composition inhibits angiogenesis in the subject. Effective below, the composition is an amount sufficient to cause cytotoxicity in the subject. In some embodiments, the secondary chemotherapeutic agent is a cytotoxic agent.

Colchicine or Thiocolchin Dimer  Compositions Containing

Colchicine or Thiocolchin Dimer

The methods described herein include the administration of a composition comprising a colchicine or thiocolchicine dimer. As used herein, "colchicine or thiocolhitine dimer" refers to a compound containing two subunits (identical or different) of colchicine, thiocolhitine or derivatives thereof. The "derivatives" of colchicine or thiocolhitine include, but are not limited to, compounds that are structurally similar to colchicine or thiocolhitine or are the same general chemical group as colchicine and thiocolhitchin. In general, derivatives or analogs of colchicine or thiocolhitchin have biological, pharmacological, chemical and / or physical properties (eg, including functionality) similar to colchicine or thiocolhitchin. In some embodiments, the colchicine or thiocolchicine dimers have one or more thiocolchicine subunits. In some embodiments, colchicine or thiocolhitine dimers comprise two thiocolhitine subunits (hereinafter referred to as "thiocolhitine dimers"). In some embodiments, colchicine or thiocolhitine dimers comprise two colchicine subunits (hereinafter referred to as "colchicine dimers").

In some embodiments, the colchicine or thiocolchicine dimer is a compound of formula (I)

<Formula I>

Wherein B in each subunit is a methoxy or methylthio group, R ₂ is methoxy, hydroxyl or methylenedioxy with R ₃ and R ₃ is methoxy, hydroxyl or together with R ₂ Methylenedioxy and X is a linking group.

A wide range of crosslinking groups can be used to introduce the linking group X. One skilled in the art recognizes that the colchicine or thiocolchicine monomer component of the dimer has a single reactive amino group, and any other reactive (nucleophilic) groups present on the intermediate can be readily protected using groups known in the art. something to do. Examples of protecting groups are described, for example, in Greene, TW, and PGM Wuts, Protective Groups in Organic Synthesis , 3rd edition, Wiley: New York, 1999, the contents of which are incorporated herein by reference in their entirety. Thus, a wide range of crosslinking groups reactive with amine functional groups can be used.

In some embodiments, the linking group X includes one or more carbon atoms. For example, commercially available (Sigma Aldrich) reagent malonyl chloride, Cl-C (O) -CH ₂ -C (O) -Cl, can be used to form colchicine dimers in which the X group is -CH _2- . . Similarly, different diacyl chlorides of various lengths can be used to form X groups of the desired length. For example, in the formula (II) where n = 8 and Y is CH ₂ , dodecanedioyl dichloride, Cl-C (O)-(CH ₂ ) ₁₀ -C (O), a commercially available (Sigma Aldrich) reagent -Cl can be used to synthesize dimers where the X group is-(CH ₂ ) _10- . For groups where Y is NH and n = 1, Reagent 3-isocyanatopropaneoyl chloride (Organic Syntheses, Coll. Vol. 6, p.715 (1988); Vol. 59, ρ. 195 (1979) ]) Can be used to synthesize a linker X wherein X is -NH-CH ₂ CH _2- . Other crosslinking reagents can be used to make the X linker. One skilled in the art will review, Wong, Shan S., Chemistry of Protein Conjugation and Cross Linking , CRC Press: Boca Raton, 1991], in particular, chapter 2, section IV (B), pages 33 to 38 (relative to amino group reactive agents); See chapter 4, section II, pages 75-103 (relative to amino group reactive crosslinkers) and chapter 7, pages 209-220 (analytical procedures and procedures for crosslinking reactions for reagents and reagents suitable for crosslinking of amino-containing compounds). . The entire contents of the above-mentioned Wong's documents, especially those specifically listed, are incorporated herein by reference.

In some embodiments, the colchicine or thiocolchicine dimer is a compound of formula (II)

<Formula II>

Wherein B ₁ is a methoxy or methylthio group, B ₂ is a methoxy or methylthio group, n is an integer from 0 to 8, Y is a CH ₂ group or when n is 1 it is also a formula NH It may be a flag.

In some embodiments, n is any one of 0, 1, 2, 3, 4, 5, 6, 7, or 8 (in some embodiments selected from the group consisting of these). In some embodiments n is 1. In some embodiments, n is 1 and Y is NH. In some embodiments n is 2.

In some embodiments, B ₁ and B ₂ are both methoxy groups. In some embodiments, B ₁ and B ₂ are both methylthio groups. In some embodiments, B ₁ is a methoxy group and B ₂ is a methyl thi group. In some embodiments, B ₁ is a methylthio group and B ₂ is a methoxy group. In some embodiments, the colchicine or thiocolchicine dimer is any of IDN5404, IDN5676, IDN5800 and IDN5801 (in some embodiments selected from the group consisting of these).

In some embodiments, the compound is thiocolchicine dimer IDN5404. IDN5404 is a compound of formula (III)

In some embodiments, the compound is thiocolchicine dimer IDN5676. IDN5676 is a compound of formula (IV)

Biocompatible polymers and carrier  protein

In some embodiments, the colchicine or thiocolhitine dimer compositions described herein further comprise a biocompatible polymer, such as a carrier protein.

The term "biocompatibility" as used herein describes a substance that does not significantly alter the biological system into which it is introduced or does not affect in an adverse manner. Biocompatible polymers include natural or synthetic biocompatible materials such as proteins, polynucleotides, polysaccharides (eg, starch, cellulose, dextran, alginate, chitosan, pectin, hyaluronic acid, etc.) and lipids. Suitable biocompatible polymers are, for example, natural or synthetic proteins such as albumin, insulin, hemoglobin, lysozyme, immunoglobulins, α-2-macroglobulin, fibronectin, vitronectin, fibrinogen, casein, etc., as well as combinations of any two or more thereof. It includes. Synthetic polymers are for example polyalkylene glycols (e.g., straight or branched chains), polyvinyl alcohols, polyacrylates, polyhydroxyethyl methacrylates, polyacrylic acids, polyethyloxazolines, polyacrylamides, poly Isopropyl acrylamide, polyvinylpyrrolidone, polylactide / glycolide, and the like and combinations thereof.

The term “protein” refers to a polypeptide or polymer (including full length or fragment) of amino acids of any length, which may be straight or branched, and may include modified amino acids and / or sandwich non-amino acids. The term also encompasses amino acid polymers that are naturally or mediatedly modified, including, for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation or any other manipulation or modification. Also encompassed by this term are polypeptides containing, for example, one or more analogs of amino acids (including, for example, non-natural amino acids, etc.) as well as other modifications known in the art. The proteins described herein may be naturally occurring, that is to say obtained or derived from natural sources (eg, blood), or may be synthesized (eg, chemically synthesized or synthesized by recombinant DNA techniques).

Examples of suitable carrier proteins include proteins commonly found in blood or plasma, including but not limited to albumin, immunoglobulins including IgA, lipoproteins, apolipoprotein B, α-acid glycoprotein, β-2- Macroglobulin, tyroglobulin, transferrin, fibronectin, factor VII, factor VIII, factor IX, factor X, and the like. In some embodiments, the carrier protein is a non-blood protein such as casein, α-lactalbumin and β-lactoglobulin. Carrier proteins may be naturally occurring or may be prepared synthetically. In some embodiments, the carrier protein is albumin, such as human serum albumin. It is human serum albumin (HSA) consists of a 585 amino acid protein and high soluble globular of M _r 65K. HSA is the most abundant protein in plasma and amounts to 70-80% of colloidal osmotic pressure in human plasma. The amino acid sequence of HSA contains a total of 17 disulfide bridges, one free thiol (Cys 34) and one tryptophan (Trp 214). Intravenous use of HSA solution may be combined with exchange transfusion in the treatment of neonatal hyperbiliruemia (see, eg, Finlayson, Seminars). in Thrombosis and Hemostasis , 6, 85-120 (1980)), for the prevention and treatment of low blood shock (see, eg, Tulis, JAMA , 237, 355-360, 460-463 (1977)) and House et al, Surgery , Gynecology and Obstetrics , 150, 811-816 (1980)]. Other albumin, such as bovine serum albumin, are also contemplated. The use of such non-human albumin may be appropriate, for example, with regard to the use of the composition in non-human mammals such as livestock animals (including domestic pets and agricultural animals).

Human serum albumin (HSA) has a large number of hydrophobic binding sites (8 total for fatty acids, endogenous ligands of HSA) and binds to a diverse set of drugs, particularly neutral and negatively charged hydrophobic compounds (Goodman et al, The Pharmacological Basis of Therapeutics , 9th edition, McGraw-Hill New York (1996)]. Two high affinity binding sites are proposed in subdomains IIA and IIIA of HSA, which are very extended hydrophobic pockets with charged lysine and arginine residues near the surface that serve as attachment points for polar ligand characteristics (eg For literature [Fehske et al, Biochem. Pharmcol . 30, 687-92 (1981)], [Vorum, Dan. Med. Bull. 46, 379-99 (1999), Kragh-Hansen, Dan.Med.Bull. 1441 , 131-40 (1990), Curry et al, Nat . Struct . Biol ., 5, 827-35 (1998), Sugio et al, Protein . Eng . , 12, 439-46, (1999). ], He et al, Nature , 358, 209-15 (1992), and Carter et al, Adv . Protein . Chem ., 45, 153-203 (1994).

To provide an example, the carrier protein is further described below. This description is understood to apply generally to biocompatible polymers.

Carrier proteins (such as albumin) in the composition generally serve as carriers for colchicine or thio colchicine dimers, that is, the carrier proteins in the composition are more effective in the aqueous medium than colchicine or thiocolchicine dimers compared to compositions that do not comprise carrier proteins. Helps to suspend or maintain suspension more easily. This may avoid the use of toxic solvents to solubilize colchicine or thiocolhitine dimers and thus reduce one or more side effects caused by the toxic solvent. In some embodiments, the composition is substantially free of surfactant, that is, when the composition is administered to a subject, the amount of surfactant in the composition is not sufficient to cause one or more side effects in the subject. In some embodiments, the composition does not contain a surfactant.

In some embodiments, the carrier protein is associated with colchicine or thiocolchicine dimer, that is to say that the composition comprises a carrier protein associated colchicine or thiocolchicine dimer. As used herein, "association" or "associated" is used in the general sense and refers to a carrier protein that affects the behavior and / or properties of colchicine or thiocolhitine dimers in aqueous compositions. For example, the carrier protein and colchicine or thiocolchicine dimer are those that are “associated” if the carrier protein makes the colchicine or thiocolchicine dimer more readily suspended in an aqueous medium, as compared to a composition without the carrier protein. Is considered. As another example, if the carrier protein stabilizes colchicine or thiocolchicine dimers in an aqueous suspension, the carrier protein and colchicine or thiocolchicine dimers are associated. For example, the carrier protein and colchicine or thiocolhitine dimer can exist as particles or nanoparticles, which are further described herein.

Colchicine or thiocolhitine dimers can be used for long periods of time, for example at least about 0.1, 0.2, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, If it remains suspended in an aqueous medium for 36, 48, 60 or 72 hours (eg, without visible precipitation or sedimentation) it is "stabilized" by the carrier protein in an aqueous suspension. Suspensions are generally suitable, but not necessarily, for administration to an individual (such as a human). The stability of the suspension is generally assessed at storage temperatures (eg room temperature (eg 20-25 ° C.) or cooling conditions (eg 4 ° C.)). For example, when observed visually at about 15 minutes after preparation of the suspension or under an optical microscope (or other suitable particle characterization technique) at 1000 magnification, the suspension is stable at storage temperature if it does not show clot or particle aggregation. It is also possible to assess stability under accelerated test conditions, for example at temperatures above about 40 ° C.

The carrier protein and colchicine or thiocolhitine dimer can be associated in various ways in the composition. For example, in some embodiments, the carrier protein can be mixed with colchicine or thiocolchicine dimers. In some embodiments, the carrier protein encapsulates or captures colchicine or thiocolchicine dimer. In some embodiments, the carrier protein is bound (eg, non-covalent) to colchicine or thiocolchicine dimers. In some embodiments, the composition may exhibit one or more of these aspects.

In some embodiments, the composition comprises particles (such as microparticles or nanoparticles) comprising essentially colchicine or thiocolchicine dimers and carrier proteins (such as albumin) in various embodiments. In some embodiments, colchicine or thiocolhitine dimers are coated with a carrier protein (such as albumin). In some embodiments, the composition comprises nanoparticles (ie, particles having an average diameter of about 1000 nanometers (nm) or less), such as averages of 900, 800, 700, 600, 500, 400, 300, 200, and 100 nm or less. It includes particles with a diameter. In some embodiments, the average diameter of the nanoparticles in the composition is about 200 nm or less. In some embodiments, the average diameter of the nanoparticles in the composition is about 20 to about 400 nm. In some embodiments, the average diameter of the nanoparticles is about 40 to about 200 nm or less. In some embodiments, the particles are sterile-filterable.

The particles (such as microparticles or nanoparticles) described herein may be present in a dry formulation (such as a lyophilized composition) or suspended in a biocompatible medium. Suitable biocompatible media include but are not limited to water, buffered aqueous media, saline, buffered saline, optionally buffered amino acid solutions, optionally buffered protein solutions, optionally buffered sugar solutions, optionally buffered vitamin solutions, optionally buffered synthetic polymer solutions , Lipid-containing emulsions and the like.

The amount of carrier protein in the compositions described herein will vary depending on the agents and other ingredients in the composition. In some embodiments, the composition comprises a carrier protein in an amount sufficient to stabilize the colchicine or thiocolchicine dimer in an aqueous suspension, eg, a stable colloidal suspension (such as a stable suspension of microparticles or nanoparticles). In some embodiments, the carrier protein is present in an amount that reduces the sedimentation rate of colchicine or thiocolchicine dimer in an aqueous medium. For particle-containing compositions, the amount of carrier protein also depends on the particle size and density of colchicine or thiocolchicine dimers.

In some embodiments, the carrier protein is present in an amount sufficient to stabilize the colchicine or thiocolchicine dimer at a particular concentration in the aqueous suspension. For example, the concentration of colchicine or thiocolchicine dimer in the composition can be, for example, about 0.01 to about 50 mg / ml, about 0.1 to about 50 mg / ml, about 1 to about 10 mg / ml, about 2 to about From about 0.01 to about 100 mg / ml, including any of 8 mg / ml, about 4 to about 6 mg / ml, and about 5 mg / ml. In some embodiments, the concentration of colchicine or thiocolchicine dimer is at least about 0.01 mg / ml, 0.03 mg / ml, 0.05 mg / ml, 0.08 mg / ml, 0.1 mg / ml, 0.3 mg / ml, 0.5 mg / ml , 0.8 mg / ml, 1 mg / ml, 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml , 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml and 50 mg / ml. In some embodiments, the carrier protein is present in an amount that avoids the use of a surfactant, such that the composition is substantially free of surfactant.

In some embodiments, the composition in liquid form may contain about 0.1% to about 50% (w / v) (eg, about 0.5% (w / v), about 5% (w / v), about 10% (w) / v), about 15% (w / v), about 20% (w / v), about 30% (w / v), about 40% (w / v) or about 50% (w / v)) Carrier proteins. In some embodiments, the composition in liquid form comprises about 0.5% to about 5% (w / v) carrier protein.

In some embodiments, the weight ratio of carrier protein, such as albumin to colchicine or thiocol hitin dimer, is such that a sufficient amount of colchicine or thiocolhitine dimer binds to or is carried by the cell. In some embodiments, the weight ratio (w / w) of the carrier protein, such as albumin to colchicine or thiocolhitine dimer, is about 0.01: 1 to about 100: 1, about 0.02: 1 to about 50: 1, about 0.05: 1 to about 20: 1, about 0.1: 1 to about 20: 1, about 1: 1 to about 18: 1, about 2: 1 to about 15: 1, about 3: 1 to about 12: 1, about 4: 1 to about 10: 1, about 5: 1 to about 9: 1, and about 9: 1. In some embodiments, the weight ratio of carrier protein (such as albumin) to colchicine or thiocolchicine dimer is about 18: 1 or less, about 15: 1 or less, 14: 1 or less, 13: 1 or less, 12: 1 or less, 11: 1 or less, 10: 1 or less, 9: 1 or less, 8: 1 or less, 7: 1 or less, 6: 1 or less, 5: 1 or less, 4: 1 or less and 3: 1 or less.

In some embodiments, the composition comprises particles (eg, microparticles or nanoparticles) comprising thiocolchicine dimers (eg, IDN5404 or IDN5676) and albumin (consisting essentially of them in various embodiments). The average diameter of the particles (eg, microparticles or nanoparticles) is about 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 micrometer or less. In some embodiments, the particles are nanoparticles, ie, particles less than about 1000 nanometers (nm). For example, the nanoparticles can be no more than 900, 800, 700, 600, 500, 400, 300, 200 and 100 nm. In some embodiments, the average diameter of the nanoparticles is about 200 nm or less. In some embodiments, the average diameter of the nanoparticles is about 20 to about 400 nm. In some embodiments, the average diameter of the nanoparticles is about 40 to about 200 nm. In some embodiments, the particles are sterile-filterable.

In some embodiments, thiocolchicine dimers (eg, IDN5404 or IDN5676) are coated with albumin. In some embodiments, the weight ratio (w / w) of albumin to colchicine or thiocolchicine dimer (eg, IDN5404 or IDN5676) is about 0.01: 1 to about 100: 1, about 0.02: 1 to about 50: 1, about 0.05 : 1 to about 20: 1, about 0.1: 1 to about 20: 1, about 1: 1 to about 18: 1, about 2: 1 to about 15: 1, about 3: 1 to about 12: 1, about 4 : 1 to about 10: 1, about 5: 1 to about 9: 1, and about 9: 1. In some embodiments, the weight ratio of albumin to colchicine or thiocolchicine dimer can be about 18: 1 or less, about 15: 1 or less, 14: 1 or less, 13: 1 or less, 12: 1 or less, 11: 1 or less, 10: 1 or less, 9: 1 or less, 8: 1 or less, 7: 1 or less, 6: 1 or less, 5: 1 or less, 4: 1 or less and 3: 1 or less.

In some embodiments, particles (eg, microparticles or nanoparticles) comprising thiocolchicine dimers (eg, IDN5404 or IDN5676) and albumin are suspended in an aqueous medium (eg, an aqueous medium containing albumin). For example, the composition may be a colloidal suspension of thiocolchicine dimers (eg, IDN5404 or IDN5676) containing particles (eg, microparticles or nanoparticles). In some embodiments, the composition is a dry composition (eg, lyophilized composition) that can be reconstituted with an aqueous suspension of thiocolchicine dimer (eg, IDN5404 or IDN5676) containing particles. In some embodiments, the concentration of thiocolchicine dimer (eg, IDN5404 or IDN5676) in the composition is, for example, about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / from about 0.1 to about 100 mg / ml including ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml and about 5 mg / ml. In some embodiments, the concentration of IDN5404 or IDN5676 is about 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / any of ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml and 50 mg / ml to be.

In some embodiments, the composition comprises a nanoparticle formulation of IDN5404 or IDN5676 (also referred to as Nab-5404 or Nab-5676 hereinafter). Nab-IDN5404 and Nab-5676 are nanoparticle formulations that stabilize IDN5404 and IDN5676 with human serum albumin, respectively. These nanoparticle formulations can be made by the methods described in US Pat. No. 5,916,596 and US Patent Application Publication 2005/0004002. When suspended in a suitable aqueous medium, such as a 0.9% sodium chloride injection or a 5% dextrose injection, Nab-5404 (or Nab-5676) forms a stable colloidal suspension of thiocolchicine dimer. The size (ie, average diameter) of the particles in the colloidal suspension may range from 20 nm to 8 micrometers, preferably from about 20 to 400 nm. Since HSA is freely soluble in water, Nab-5404 (or Nab-5676) can be used in dilute concentrations (0.1 mg / ml IDN5404 or IDN5676, including, for example, about 2 mg / ml to about 8 mg / ml, about 5 mg / ml). ) To a wide range of concentrations (from 20 mg / ml IDN5404 or IDN5676). In some embodiments, the concentration of IDN5404 or IDN5676 is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and Which is 20 mg / ml.

Pharmaceutical compositions, unit dosages, and Kit

Also provided are pharmaceutical compositions comprising colchicine or thiocolhitine dimers. Pharmaceutical compositions may be suitable for the various modes of administration described herein, including, for example, systemic or topical administration. The pharmaceutical composition may be in the form of eye drops, injectable solutions, or in a form suitable for inhalation (through the mouth or nose) or oral administration. The pharmaceutical compositions described herein may be packaged in a single unit dosage form or in multiple dosage forms. In some embodiments, the composition is suitable for administration to a human. In some embodiments, the composition is suitable for administration to a mammal, such as a domestic pet and an agricultural animal, in a veterinary context. Numerous suitable composition formulations exist (see, eg, US Pat. Nos. 5,916,596 and 6,096,331).

In some embodiments, the pharmaceutical composition comprises colchicine or thiocolchicine dimers, carrier proteins, and pharmaceutically acceptable carriers suitable for intraocular injection. In some embodiments, the pharmaceutical composition comprises colchicine or thiocolchicine dimers, carrier proteins, and pharmaceutically acceptable carriers suitable for topical application to the eye. In some embodiments, the pharmaceutical composition comprises colchicine or thiocolchicine dimers, carrier proteins, and pharmaceutically acceptable carriers suitable for intraarterial injection.

Pharmaceutical compositions are generally formulated into sterile and substantially isotonic compositions. For injection, the pharmaceutical composition may be in the form of a physiologically compatible buffer such as a liquid solution, for example Hank's solution or Ringer's solution. Pharmaceutical compositions may also be in solid form and redissolved or resuspended immediately prior to use. Lyophilized compositions are also included.

For oral administration, the pharmaceutical composition may contain pharmaceutically acceptable excipients such as binders (eg, pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); Fillers (eg lactose, microcrystalline cellulose or calcium hydrogen phosphate); Lubricants (eg magnesium stearate, talc or silica); Disintegrants (eg potato starch or sodium starch glycolate); Or wetting agents (eg sodium lauryl sulfate), for example in the form of tablets or capsules in conventional manner. Liquid preparations for oral administration may be, for example, in the form of solutions, syrups or suspensions, or may be present as a dry product composed by water or other suitable vehicle before use. Such liquid formulations include pharmaceutically acceptable additives such as suspending agents (eg, sorbitol syrup, cellulose derivatives or hydrogenated edible fats); Emulsifiers (eg lecithin or acacia); Non-aqueous vehicles (eg, ationd oils, oily esters, ethyl alcohol or fractionated vegetable oils); And preservatives (eg, methyl or propyl-p-hydroxybenzoate or sorbic acid). The formulations may also contain buffer salts, flavoring agents, coloring agents and sweetening agents as necessary.

The present invention provides, in some embodiments, a composition comprising a colchicine or thiocolchicine dimer, a carrier protein, and a pharmaceutically acceptable carrier suitable for administration to the eye. Such pharmaceutical carriers can be sterile liquids such as water and oils such as those of petroleum, animal, plant or synthetic origin such as peanut oil, soybean oil, mineral oil and the like. Saline solutions and aqueous dextrose, polyethylene glycol (PEG) and glycerol solutions can also be used as liquid carriers, especially for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, sodium stearate, glycerol monostearate, glycerol, propylene, water and the like. If desired, the pharmaceutical composition may also contain small amounts of wetting or emulsifying agents, or pH buffers. The components of the composition can be trapped in a polymer or fibrin glue to control the release of molecules. Such compositions may be in the form of solutions, suspensions, emulsions, ointments, gels, or other solid or semisolid compositions and the like. The composition typically has a pH of 4.5 to 8.0. The composition should also be formulated to have an osmotic value compatible with the waterproofing of the eye and eye tissue. Such osmotic values are generally in the range of about 200 to about 400 milliomols of water / kg of water (“mOsm / kg”), preferably about 300 mOsm / kg.

In some embodiments, the composition is formulated into a pharmaceutical composition that is applied for intravenous, intraperitoneal, or intravitreal injection according to conventional procedures. Typically, the injectable composition is a solution of isotonic sterile aqueous buffer. If desired, the composition may also include a local anesthetic such as lignocaine and a solubilizer to relieve pain at the injection site. Generally, as a lyophilized powder or anhydrous concentrate in a hermetically sealed container such as a sachet or ampoule indicating the amount of active agent, the components are supplied separately or mixed together in a unit dosage form. When the composition is injected and administered, it is administered in an infusion bottle containing sterile pharmaceutical grade water or saline. When the composition is administered by injection, an ampoule of sterile water or saline for injection can be provided so that the components can be mixed before administration.

The composition may further comprise additional ingredients such as preservatives, buffers, osmotic modifiers, antioxidants and stabilizers, nonionic wetting or degreasing agents, viscosity increasing agents and the like.

Preservatives suitable for use in the solution include polyquaternium-1, benzalkonium chloride, thimerosal, chlorobutanol, methyl paraben, propyl paraben, phenylethyl alcohol, disodium disodium, sorbic acid, benzethonium chloride, and the like. do. Typically (but not required) such preservatives are used at levels of 0.001% to 1.0% by weight.

Suitable buffers include boric acid, sodium and potassium bicarbonate, sodium and potassium borate, sodium and potassium carbonate, sodium acetate, sodium diphosphate, etc., to maintain pH at about pH 6 to pH 8, preferably at about pH 7 to pH 7.5. Include in sufficient quantity.

Suitable osmotic modulators are dextran 40, dextran 70, dextrose, glycerin, potassium chloride, propylene glycol, sodium chloride and the like, with sodium chloride equivalents in the ophthalmic solution in the range of 0.9 ± 0.2%.

Suitable antioxidants and stabilizers include sodium bisulfite, sodium metabisulfite, sodium thiosulfite, thiourea, and the like. Suitable humectants and detergents include polysorbate 80, polysorbate 20, poloxamer 282 and tyloxapol. Suitable viscosity increasing agents are dextran 40, dextran 70, gelatin, glycerin, hydroxyethylcellulose, hydroxymethylpropylcellulose, lanolin, methylcellulose, petrolatum, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, carboxy Methylcellulose and the like.

Using a viscosity increasing agent to provide a topical composition with a viscosity that is higher than the viscosity of a simple aqueous solution may be desirable to increase the absorption of the active compound into the eye by the target tissue or to increase the retention time in the eye. Such viscosity enhancers are, for example, polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxy propyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, hydroxy propyl cellulose or other agents known in the art. It includes. Such agents are typically used at levels of 0.01% to 2% by weight.

The compositions described herein also include other agents, excipients, or stabilizers to enhance the properties of the composition. For example, to increase stability by increasing the negative zeta potential of the nanoparticles, certain negatively charged components can be added. The negatively charged components include glycolic acid, cholic acid, kenodeoxycholic acid, taurocholic acid, glycokenodeoxycholic acid, taurokenodeoxycholic acid, lithocholic acid, ursodeoxycholic acid, dehydrocholic acid and other bile acids. Bile salts; Phosphatidylcholine, including palmitoyl oleoylphosphatidylcholine (including phosphatidylcholine, including phosphatidylcholine) Phospholipids including, but not limited to, the base phospholipids. Other phospholipids include L-α-dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), and other related compounds. Negatively charged surfactants or emulsifiers such as sodium cholesteryl sulfate and the like are also suitable as additives.

In addition, unit dosage forms of colchicine or thiocolchicine dimer compositions, such as colchicine or thiocolchicine dimers, for example from about 0.2 mg to about 50 mg, about 0.5 mg to about 30 mg, about 1 mg to about 20 mg Or, unit dosages containing from about 0.1 mg to about 50 mg, including about 15 mg, are provided. The term “unit dosage form” refers to physically distinct units suitable for single administration to an individual, each unit comprising a predetermined amount of active substance calculated with the appropriate pharmaceutical carrier, diluent, or excipient to achieve the desired therapeutic effect. It contains. These unit dosage forms may be stored in a suitable package of single or multiple unit dosage forms and may also be further sterilized and sealed. In some embodiments, unit dosage forms of colchicine or thiocolchicine dimers and carrier proteins suitable for intraocular injection are provided. In some embodiments, unit dosage forms of colchicine or thiocolchicine dimers and carrier proteins suitable for topical application to the eye are provided. In some embodiments, unit dosage forms of colchicine or thiocolchicine dimers and carrier proteins suitable for intraarterial injection are provided.

Also provided are products comprising the compositions described herein in suitable packaging. Suitable packaging for the compositions (eg ophthalmic compositions) described herein are known in the art and include, for example, vials (eg, sealed vials), tubes, ampoules, bottles, jars, flexible packaging (eg, Sealed Mylar or plastic bags) and the like. These products may be further sterilized and / or sealed.

The invention also provides kits (or unit dosage forms and / or products) comprising the compositions described herein, and may further comprise instructions regarding how to use the compositions, such as the uses described herein. Kits described herein may further include other materials desirable from a commercial and user standpoint, including other buffer diluents, filters, needles, syringes, and package inserts, with instructions for performing any of the methods described herein. For example, in some embodiments, the kit includes one or more buffers, diluents, filters, needles, with colchicine or thiocolchicine dimers, carrier proteins, pharmaceutically acceptable carriers suitable for intraocular injections, and instructions for performing intraocular injections. , Syringe and package insert. In some embodiments, the pharmaceutical composition comprises one or more buffers, diluents, filters, needles, with colchicine or thiocolchicine dimers, carrier proteins, pharmaceutically acceptable carriers suitable for intraarterial injection, and instructions for performing intraarterial injection. Syringe and package insert.

While the invention has been described in some detail above by way of explanation and illustration for clarity of understanding, it will be apparent to those skilled in the art that minor changes and modifications may be made. Accordingly, the description and examples should not be understood as limiting the scope of the invention.

Example  One. Nab -5404 and Nab -5676 In vitro  Cytotoxic activity

The cytotoxic activity of Nab-5404 and Nab-5676 was evaluated in vitro using MX-1 breast carcinoma, HepG2 liver cancer, HT-29 colon carcinoma cell line and normal primary rat hepatocytes. Nab-5404 and Nab-5676 were exposed to cells at increasing concentrations at 37 ° C. for 72 hours. Survival was analyzed using a Cell Titer Blue cell viability analyzer (Promega, Madison, Wis.). IC50 was calculated with a one-phase exponential decay equation using Prism software (GraphPad, San Diego, Calif.).

Nab-5404 showed cytotoxic activity against HepG2 hepatocarcinoma cells and primary rat hepatocytes with IC50 of 16 and 9 μg / ml, respectively. Nab-5404 and Nab-5676 were moderate cytotoxic to MX-1 cells with IC 50 of 43 and 54 μg / ml, respectively. Nab-5404 and Nab-5676 were low in activity with IC50 of 110 and 149 μg / ml for HT-29 cells, respectively. Nab-5676 showed little or no cytotoxic activity in HepG2 cells or primary rat hepatocytes. The results are summarized in Table 1.

Cell line Nab-5404 IC50 (μg / ml) Nab-5676 IC50 (μg / ml) MX-1 Breast Carcinoma 43 54 HepG2 Liver Cancer 16 125 HT-29 Colon Carcinoma 110 149 Primary Rat Hepatocytes 9 Inert

Example  2. Nab -5404 and Nab -5676 Antimicrobial  activation

The microtubule depolymerization activity of Nab-5404 and Nab-5676 was tested using the MX-1 breast carcinoma cell line. MX-1 cells were seeded on cover slips and treated with Nab-5404 or Nab-5676 for 2 hours at 37 ° C. over a concentration range of 0.01-100 μg / ml. After incubation, cells were fixed and stained for tubulin and actin. Tubulin was stained with monoclonal anti-tubulin antibody and actin was stained with fluorescein-labeled paloidine. Microtubule networks were visualized and analyzed using ImagePro software (MediaCybernetics, Inc., Silver Spring MD). Prism software (GraphPad, San Diego, Calif.) Was used to calculate IC 50 of Nab-5404 and Nab-5676 activity against microtubule attenuation.

Nab-5404 and Nab-5676 exhibit potent microtubule depolymerization activity with IC50 calculated values of 0.06 μg / ml and 0.12 μg / ml, respectively (FIGS. 1A and 1B). Even at the lowest concentration tested 0.6 μg / ml, the microtubule network was completely destroyed 2 hours after incubation with Nab-5404 or Nab-5676 (FIGS. 2A-2C). In contrast, actin bundles were not affected by Nab-5404 or Nab-5676 at all drug concentrations.

Example  3. Nab -5404 and Nab -5676 Antiangiogenesis  activation

To study the effects of Nab-5404 and Nab-5676 on angiogenesis, compounds were evaluated in microangiogenesis assays using the TCS Cellwork Angiokit Model (TCS CellWorks Ltd., Botolph Claydon, Buckingham UK). Angiokitt models use human endothelial cells co-cultured with other human cells. Endothelial cells initially formed small islands in the culture substrate and then started to proliferate, enter the mobile phase, migrate through the substrate, and form a tubular tubular structure. They gradually connected to form a tubular network very similar to capillaries (9-11 days). The tubules were stained positive for von Willebrand factor, CD31 (PECAM-I) and ICAM-2.

In addition to Nab-5404 and Nab-5676, the known vascular targeting agent combretastatin 4-phosphate (CA4P) was tested for in vitro antiangiogenic activity. TCS Cellwork Angiokit model was used according to manufacturer's instructions. As described above, human endothelial cells were co-cultured with human fibroblast cells in 24-well plates and exposed to Nab-5404, Nab-5676 or CA4P over a range of concentrations (0.01-100 μg / ml). After 11 or 12 days of incubation, cells were fixed and stained with secondary antibody conjugates, colored substrates and monoclonal antibodies to CD31 to visualize tubules. The length of the tubules was analyzed using ImagePro software and the IC50 of each compound was calculated using Prism software.

Nab-5404 and Nab-5676 both exhibited anti-angiogenic activity by inhibiting tubule formation and disrupting established tubules. In the first experiment, cells were treated with the composition on day 1 and stained on day 12 to analyze the inhibition of tubule formation. In the second experiment, cells were treated on day 8 and stained on day 11, and in the third experiment, cells were treated on day 11 and stained on day 12 to analyze the breakdown of established tubules.

Nab-5404 and Nab-5676 were both able to inhibit the formation of new microvessels (FIGS. 3 and 4). In the second experiment, both Nab-5404 and Nab-5676 were able to inhibit tubule formation and / or destroy established tubules (FIG. 5). Nab-5404 destroyed the established tubules more strongly than Nab-5676 (FIGS. 6, 7 and 9). The IC 50 calculation for Nab-5404 was 0.002 μg / ml, which was 10 times stronger than Nab-5676 with an IC50 of 0.02 μg / ml. Nab-5404 was as effective as CA4P (IC 50 = 0.003 μg / ml) in the destruction of established tubules (FIGS. 6, 8 and 9).

Example  4. Nab -5404 and Nab Anti-tumor activity of -5676

Antitumor activity of Nab-5404 and Nab-5676 was evaluated in xenograft mouse models. Compounds were tested in two cycles of the first high dose schedule followed by the second high dose schedule against established HT-29 colon cell tumors in vivo. Irinotecan was used as a positive control in the test. Mice (n = 10) were divided into eight groups and mice were administered Nab-5404 and Nab-5676 compounds by intravenous injection. In the first cycle, Nab-5404 or Nab-5676 was administered 4 times every 3 days from 0 to 14 days, and in the 2nd cycle, Nab-5404 or Nab-5676 4 times every 3 days from 15 to 30 days. Was administered. Irinotecan was administered four times every three days at a dose of 60 mg / kg using intravenous injection. The populations are shown in Table 2, with the 1st cycle representing 0 to 14 days and the 1st cycle representing 15 to 30 days.

process First Cycle Dose 1st cycle schedule Second cycle dose 2nd cycle schedule Control vehicle Irinotecan 60 mg / kg q3d x 4 Nab-5404 3.4 mg / kg q3d x 4 40 mg / kg q3d x 4 Nab-5404 2.5 mg / kg q3d x 4 30 mg / kg q3d x 4 Nab-5404 1.7 mg / kg q3d x 4 20 mg / kg q3d x 4 Nab-5676 3.4 mg / kg q3d x 4 40 mg / kg q3d x 4 Nab-5676 2.5 mg / kg q3d x 4 30 mg / kg q3d x 4 Nab-5676 1.7 mg / kg q3d x 4 20 mg / kg q3d x 4

As shown in Figures 10 and 11, at low doses both Nab-5404 and Nab-5676 significantly inhibited tumor growth (p values for 3.4, 2.5 and 1.7 mg / kg Nab-5404 were 0.02, 0.007 , 0.001 and p values for 3.4, 2.5, and 1.7 mg / kg Nab-5676 were 0.04, 0.003 and 0.0004). The antitumor activity of Nab-5404 and Nab-5676 at low doses means that these compounds have antiangiogenic activity. At higher doses, significantly higher tumor regression occurred after treatment with Nab-5404 compared to Nab-5676. Based on the weight loss data, the MTD of Nab-5404, which shows significant tumor regression in this model, was approximately 30 mg / kg.

In separate experiments combretastatin 4-phosphate (CA4P) was also administered to HT-29 tumor bearing mice. When tumors reached 900 mm ³ , mice (n = 10) were treated with CA4P or vehicle, with MTD of 100 mg / kg, on a schedule of four times daily. CA4P was found to be inactive in this tumor model without difference from the control vehicle (FIG. 12).

While the invention has been described in some detail above by way of explanation and illustration for clarity of understanding, it will be apparent to those skilled in the art that minor changes and modifications may be made. Accordingly, the description and examples should not be understood as limiting the scope of the invention.

Claims (31)

Administering to the subject a composition comprising a carrier protein and a colchicine or thiocolhitine dimer, the composition being an amount effective to inhibit angiogenesis, and the amount of the composition is insufficient to cause significant cytotoxicity in the individual. The method of inhibiting angiogenesis in a subject. A method comprising treating to a subject a composition comprising a carrier protein and a colchicine or thiocolchicine dimer, wherein the composition is an amount effective to treat a non-tumoral angiogenesis related disease. The method of claim 2, wherein the non-neoplastic angiogenesis related disease is an ocular disease. 4. The method of claim 3, wherein the ocular disease is senile macular degeneration, diabetic retinopathy or neovascular glaucoma. The method of claim 2, wherein the non-tumor angiogenesis related disease is a cardiovascular disease. The method of claim 5, wherein the cardiovascular disease is restenosis or atherosclerosis. The method of claim 2, wherein the non-tumoral angiogenesis related disease is rheumatoid arthritis or psoriasis. The method of claim 2, wherein the amount of the composition is insufficient to cause significant cytotoxicity in the subject. Administering to a subject a composition comprising a carrier protein and a colchicine or thiocolchicine dimer, the composition being an amount effective to treat cancer, the amount of the composition being insufficient to cause significant cytotoxicity in the subject. The method of treating cancer in a subject. 10. The method of claim 9, wherein the amount of colchicine or thiocolchicine dimer in the composition is less than about 15% of the corresponding MTD of colchicine or thiocolchicine dimer. The method of claim 10, wherein the cancer is a solid tumor. The method of claim 11, wherein the cancer is a metastatic solid tumor. The method according to any one of claims 1 to 12, wherein the colchicine or thiocolchicine dimer is a compound of formula (II). <Formula II>

In the formula, B ₁ is a methoxy or methylthio group; B ₂ is a methoxy or methylthio group; n is an integer from 0 to 8; Y may be a CH ₂ group or when n is 1 it may also be a group of formula NH. The method of claim 13, wherein Y is CH ₂ . The method of claim 13, wherein n is 1 and Y is NH. The method of claim 13, wherein B ₁ and B ₂ are both methylthio groups. The method of claim 13, wherein the compound is IDN5404. The method of claim 13, wherein the compound is IDN5676. 13. The method of any one of claims 1-12, wherein the amount of colchicine or thiocolchicine dimer in the composition is less than about 15 mg / m ² . The method of claim 1, wherein the composition is administered once weekly. The method of claim 20, wherein the composition is administered for at least 3 months without interruption. The method of claim 1, wherein the composition is substantially free of surfactant. The method of claim 1, wherein the carrier protein is albumin. The method of claim 23, wherein the ratio of albumin to colchicine or thiocolchicine dimer is less than about 18: 1. The method of claim 1, wherein the composition comprises a nanoparticle comprising a carrier protein and a colchicine or thiocolchicine dimer. The method of claim 25, wherein the carrier protein is albumin. 27. The method of claim 26, wherein the ratio of albumin to colchicine or thiocolchicine dimer is less than about 18: 1. The method of claim 25, wherein the average diameter of the nanoparticles is 200 nm or less. The method of claim 25, wherein the composition is substantially free of surfactant. The method of claim 1, wherein the composition is Nab-5404. The method of claim 1, wherein the subject is a human.

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